http://rdf.ncbi.nlm.nih.gov/pubchem/patent/AR-112454-A2
Outgoing Links
| Predicate | Object |
|---|---|
| assignee | http://rdf.ncbi.nlm.nih.gov/pubchem/patentassignee/MD5_5ace6b22056a2824f77aac332b85adba |
| classificationIPCInventive | http://rdf.ncbi.nlm.nih.gov/pubchem/patentipc/C12N5-00 http://rdf.ncbi.nlm.nih.gov/pubchem/patentipc/C12P21-02 |
| filingDate | 2018-07-16-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
| publicationDate | 2019-10-30-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
| publicationNumber | AR-112454-A2 |
| titleOfInvention | METHOD FOR THE PRODUCTION OF POLYPEPTIDES |
| abstract | An improved system is provided for the large-scale production of proteins and a-b in cell culture, in particular, in media characterized by one or more of the following characteristics: 1) a cumulative concentration of amino acids greater than about 70 mM; 2) a cumulative glutamine to cumulative asparagine molar ratio of less than about 2; 3) a molar ratio of cumulative glutamine to cumulative total amino acids of less than about 0.2; 4) a molar ratio of accumulated inorganic ion to accumulated total amino acids of between about 0.4 and 1; or 5) a combined accumulated glutamine and accumulated asparagine concentration of between approximately 16 and 36 mM. The use of such a system allows high levels of a-b production, and reduces the accumulation of certain undesirable factors, such as ammonium and lactate. Additionally, culture methods are provided that include a change in temperature, which usually comprises a decrease in temperature when the culture has reached about 20% - 80% of its maximum cell density. Alternatively or additionally, the present provides methods such that, after reaching a peak, lactate and ammonium levels in the culture decrease over time. Claim 1: A method for producing Etanercept in a large-scale production cell culture, characterized in that it comprises the steps of: providing a cell culture comprising; Chinese hamster ovary (CHO) cells containing a gene encoding Etanercept, where said gene is expressed under cell culture conditions; and a medium containing glutamine, which has a cumulative amount of amino acids per unit volume greater than 70 mM, a cumulative glutamine to cumulative asparagine molar ratio of less than 2, and a cumulative glutamine to cumulative amino acid molar ratio totals less than 0.2, where the total cumulative amount of histidine, isoleucine, leucine, methionine, phenylalanine, tryptophan, tyrosine, and proline per unit volume in said medium is greater than 25 mM; where the initial glutamine concentration of said medium is less than or equal to 13 mM; maintaining said culture in an initial growth phase under a first set of conditions wherein said first set of conditions comprises a temperature between 30-42 ° C; changing the temperature, so that a second set of culture conditions is applied where the second set of culture conditions comprises a second range of temperatures that is 25 to 41 ° C and where the temperature is lower than the previous temperature; maintaining said culture for a second period of time under the second set of conditions for a second period of time so that the polypeptide accumulates in the cell culture where the large-scale production cell culture is at least 500 L. |
| priorityDate | 2004-08-27-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
| type | http://data.epo.org/linked-data/def/patent/Publication |
Incoming Links
Total number of triples: 34.